Knop R H, Chen C W, Mitchell J B, Russo A, McPherson S, Cohen J S
Biochim Biophys Acta. 1985 May 30;845(2):171-7. doi: 10.1016/0167-4889(85)90174-0.
Dynamic intracellular ATP and Pi levels were measured non-invasively for Chinese hamster V79 cells by 31P-NMR under conditions of thermotolerance and heat-shock protein induction. High densities of cells were embedded in agarose strands, placed within a standard NMR sample tube, and perfused with medium maintained either at 37 or 43 degrees C at pH 7.35. Cell survival and heat-shock protein synthesis were assessed either from parallel monolayer cultures or cells dislodged from the agarose strands post-treatment. Thermotolerance (heat resistance) and heat-shock protein synthesis was induced by a 1 h exposure to 43 degrees C followed by incubation for 5 h at 37 degrees C. After the 5 h incubation at 37 degrees C, marked thermal resistance was observed in regard to survival with concomitant synthesis of two major heat-shock proteins at 70 and 103 kDa. Studies were also conducted where tolerance and heat-shock protein synthesis were partially inhibited by depletion of cellular glutathione (GSH) prior to and during heat treatment. Dynamic measurement of intracellular ATP of cells heated with or without GSH depletion revealed no change in steady-state levels immediately after heating or during the 5 h post-heating incubation at 37 degrees C where thermotolerance and heat-shock proteins develop. These data are consistent with other reported data for mammalian cells and indicate that the steady-state ATP levels in mammalian cells remain unchanged during and after the acquisition of the thermotolerant state.
在耐热性和热休克蛋白诱导条件下，通过31P-NMR对中国仓鼠V79细胞的细胞内ATP和磷酸水平进行了无创测量。将高密度细胞包埋在琼脂糖链中，置于标准NMR样品管内，并用pH值为7.35、温度维持在37或43摄氏度的培养基进行灌注。细胞存活率和热休克蛋白合成通过平行单层培养或处理后从琼脂糖链上分离的细胞进行评估。耐热性（热抗性）和热休克蛋白合成通过1小时暴露于43摄氏度，然后在37摄氏度孵育5小时来诱导。在37摄氏度孵育5小时后，观察到显著的耐热性，同时合成了两种主要的热休克蛋白，分子量分别为70和103 kDa。还进行了一些研究，在热处理之前和期间通过消耗细胞内谷胱甘肽（GSH）来部分抑制耐受性和热休克蛋白合成。对有或没有GSH消耗的加热细胞的细胞内ATP进行动态测量，结果显示在加热后立即或在37摄氏度进行5小时加热后孵育期间（此时耐热性和热休克蛋白形成），稳态水平没有变化。这些数据与其他关于哺乳动物细胞的报道数据一致，表明在获得耐热状态期间和之后，哺乳动物细胞中的稳态ATP水平保持不变。
